Team:Chiba/Calendar-Home/25 August 2008

From 2008.igem.org

>Home | Notebook

24 August 2008 <|> 26 August 2008

Contents

Laboratory work

Team:Input

The following glycerol Stock was made.

[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](insert check OK)


Picked the colony and incubate with 2mL of LB-Ampicillin Medium for 12 hours at 37 degrees. Took 180μl from culture,and mixed with 880μlLB-Amp Medium.(OD=0.485)

Liquid culture(OD=0.495) 0.67μl
60%Glycerol 0.33μl
20%Glycerol Stock 1.00ml

Team:Communication

(24/8)--->Digestion
  1. Double Digestion:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007)
  2. Double Digesiton:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)
  3. Single Digestion:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)
  4. [http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)
Sample No. 1234
Sample DNA(μL) 5511
EcoRI(μL) 0.50.5--
SpeI(μL) 0.50.5--
Buffer 1 11--
Buffer 3(μL) --1-
BSA(μL) 11--
dH2O(μL) 227.5-
TOTAL(μL) 1010101
--->Gel Check
Chiba-0825-1.JPG
Sample No. 1~34
Sample DNA(μL) 101
Loading Dye(μL) 21
dH2O(μL) -4
TOTAL(μL) 126
From right;
  1. Double Digestion:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007) -> OK
  2. Double Digesiton:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007) -> ?(low)
  3. Single Digestion:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007) -> ?(low)
  4. [http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007) -> OK(low)


--->Digestion (Double Digestion)

  1. [http://partsregistry.org/Part:BBa_C0178 BBa_C0178]
  2. [http://partsregistry.org/Part:BBa_C0170 BBa_C0170]
  3. [http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007)
Sample No. 123
Sample DNA(μL) 33ng/μl×60μl33ng/μl×60μl100ng/μl×20μl
PstI(μL) 222
XbaI(μL) 22-
SpeI(μL) --2
Buffer 2(μL) --5
Buffer 3(μL) 1010-
BSA(μL) 10105
dH2O(μL) 161616
TOTAL(μL) 10010052

Team:Output

PCR

  • [http://partsregistry.org/Part:BBa_E2030 BBa_E2030](1)
  • [http://partsregistry.org/Part:BBa_J33202 BBa_J33202](2)
  • BL21(3)
  • [http://partsregistry.org/Part:BBa_J52008 BBa_J52008](4)
  • [http://partsregistry.org/Part:BBa_I712019 BBa_I712019](5)
No. 12345
DNA tamplate(μL) 11111
FW primer(μL) 5(Venus_YFP_fwd)5(LacZ_fwd)5(LacZ_fwd)5(rLUC_fwd)5(fLuc_fwd)
VR primer(μL) 5(VR)5(LacZ_rev)5(LacZ_rev)5(VR)5(VR)
dNTPmix(μL) 1010101010
Thermo pol buffer(μL) 1010101010
DNA pol(VENT)(μL) 11111
dH2O(μL) 6868686868
TOTAL(μL) 100100100100100

-->Gel Check


No. 12345
DNA tamplate(μL) 11111
loading Dye(μL) 11111
dH2O(μL) 44444
TOTAL(μL) 66666

Mini prep

  • [http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
  • [http://partsregistry.org/Part:BBa_F2620 BBa_F2620]


Transformation

  • [http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
  • PUC19
  • pGFPUV
  • pGEX Venus YFP
  • Membren Venus YFP

PCR

  • [http://partsregistry.org/Part:BBa_J63001 BBa_J63001](1)
No. 1
DNA tamplate(μL) 1
FW primer(Venus)(μL) 5
VR primer(μL) 5
dNTPmix(μL) 10
Thermo pol buffer(μL) 10
DNA pol(VENT)(μL) 1
dH2O(μL) 68
TOTAL(μL) 100